Self-nanoemulsifying oily formulation for the administration of poorly water-soluble drugs

ABSTRACT

A pharmaceutical composition in a form of an anhydrous self-nanoemulsifying oily formulation comprising: one or more therapeutic agent(s) which have low solubility in water or are water-insoluble, vitamin E, one co-solvent selected from propylene glycol and ethanol and mixture thereof one surfactant selected from tyloxapol and from mixture of tyloxapol and TPGS, and optionally, a bioenhancer.

The present invention relates to a pharmaceutical excipient formulation,more particularly to a pharmaceutical excipient composition consistingin a self-nanoemulsifying oily formulation (SNEOF) enhancing theabsorption of poorly water soluble drugs, particularly the oralabsorption of taxoids and cytotoxic agents, based on improveddissolution and absorption of the drug; and providing a dose-AUC linearpharmacokinetic of the drug.

The clinical use of some drugs is only possible if a specific drugdelivery system is developed to transport them to their therapeutictarget in the human body. This problem is particularly critical forwater insoluble or poorly water soluble compounds for which directinjections may be impossible or problematic.

A few examples of therapeutic substances, which are poorly hydrosoluble,are the following: Palmitoyl Rhizoxin, Penclomedine, Vitamin A and itsderivatives (retinoic acid, isotretinoin, etc.), Tamoxifen, Etoposide,Campothecin, Navelbine, Valproic acid, Tacrolimus, Sirolimus(Rapamycin), Cyclosporin A, Clarithromicin, Testosterone, Estradiol,Progesterone, Ciprofloxacine, Fenofibrate, Benzafibrate, Azithromicine,Itraconazole, Miconazole, Propofol, Brimonidine, Latanoprost, andPaclitaxel.

Paclitaxel, one of the best known taxoids, disrupts tubulin dynamics. Ithas a significant clinical activity against a broad range of tumor typesincluding breast, lung, head and neck, bladder, and platinum-refractoryovarian carcinoma (E. K. Rowinsky. The development and clinical utilityof the taxoid class of antimicrotubule chemotherapy agents. Annu RevMed. 48: 353-74 (1997)). However, paclitaxel has a low therapeuticindex. It is a complex diterpenoid product, with a bulky, extended fusedring system as well as a number of hydrophobic substituents, which leadto its poor solubility in water (1 μg/ml) 30 resulting in seriousformulation problems (R. T. Liggins, W. L. Hunter, H. M. Burt.Solid-state characterization of paclitaxel. J Pharm Sci. 86: 1458-63(1997)). It is highly lyophobic and the solubility of paclitaxel inlipophilic solvents, such as soybean oil is quite low and precludes theuse of simple oil-in-water emulsions for formulation considerations. Thecommercially available product, Taxol®, is currently formulated forsystemic administration in a mixture of ethanol and polyoxyethylatedcastor oil (Cremophor EL); the latter appears to be primarilyresponsible for drug related hypersensitivity reactions, rather than thedrug itself (R. E. Gregory, A. F. De Lisa. Paclitaxel: a newantineoplastic agent for refractory ovarian cancer. Clin Pharm. 12:401-15 (1993)). Moreover, polyoxyethylated castor oil also causes thenonlinear pharmacokinetic behavior of paclitaxel (A. Sparreboom, O. vanTellingen, W. J. Nooijen, J. H. Beijnen. Nonlinear pharmacokinetics ofpaclitaxel in mice results from the pharmaceutical vehicle Cremophor EL.Cancer Res. 56: 2112-5 (1996); O. van Tellingen, M. T. Huizing, V. R.Panday, J. H. Schellens, W. J. Nooijen, J. H. Beijnen. Cremophor ELcauses (pseudo-) non-linear pharmacokinetics of paclitaxel in patients.Br J. Cancer 81: 330-5 (1999)).

The current approaches for reducing the side effects of the actualcommercial product are mainly focused on developing formulations thatare devoid of polyoxyethylated castor oil. Several attempts have beenmade to deliver paclitaxel using alternative systems, such asnanoparticles (R. Cavalli, O. Caputo, M. R. Gasco. Preparation andcharacterization of solid lipid nanospheres containing paclitaxel. Eur JPharm Sci. 10: 305-9 (2000); S. S. Feng, G. F. Huang, L. Mu. Nanospheresof biodegradable polymers: a system for clinical administration of ananticancer drug paclitaxel (Taxol). [In Process Citation]. Ann Acad MedSingapore. 29: 633-9 (2000)), liposomes (P. Crosasso, M. Ceruti, P.Brusa, S. Arpicco, F. Dosio, L. Cattel. Preparation, characterizationand properties of sterically stabilized paclitaxel-containing liposomes.J. Controlled Release. 63: 19-30 (2000); A. Sharma, R. M. Straubinger.Novel taxol formulations: preparation and characterization oftaxol-containing liposomes. Pharm Res. 11: 889-96 (1994)), water-solubleprodrugs (J. M. Terwogt, B. Nuijen, W. W. T. B. Huinink, J. H. Beijnen.Alternative formulations of paclitaxel. Cancer Treat Rev. 23: 87-95(1997); A. Pendri, C. D. Conover, R. B. Greenwald. Antitumor activity ofpaclitaxel-2′-glycinate conjugated to poly(ethylene glycol): awater-soluble prodrug. Anticancer Drug Des. 13: 387-95 (1998)),emulsions (P. P. Constantinides, K. J. Lambert, A. K. Tustian, B.Schneider, S. Lalji, W. Ma, B. Wentzel, D. Kessler, D. Worah, and S. C.Quay. Formulation development and antitumor activity of afilter-sterilizable emulsion of paclitaxel. Pharm Res. 17: 175-82(2000); B. B. Lundberg. A submicron lipid emulsion coated withamphipathic polyethylene glycol for parenteral administration ofpaclitaxel (Taxol®). J. Pharm Pharmacol. 49: 16-21 (1997); P. Kan, Z. B.Chen, C. J. Lee, I. M. Chu. Development of nonionicsurfactant/phospholipid o/w emulsion as a paclitaxel delivery system. JControlled Release. 58: 271-8 (1999), P. Simamora, R. M. Dannenfelser,S. E. Tabibi, S. H. Yalkowsky. Emulsion formulations for intravenousadministration of paclitaxel. PDA J Pharm Sci Technol. 52: 170-2 (1998))and microspheres (R. T. Liggins, S. D'Amours, J. S. Demetrick, L. S.Machan, H. M. Burt. Paclitaxel loaded poly(L-lactic acid) microspheresfor the prevention of intraperitoneal carcinomatosis after a surgicalrepair and tumor cell spill [In Process Citation]. Biomaterials. 21:1959-69 (2000); Y. M. Wang, H. Sato, I. Adachi, I. Horikoshi.Preparation and characterization of poly(lactic-co-glycolic acid)microspheres for targeted delivery of a novel anticancer agent, taxol.Chem Pharm Bull (Tokyo). 44: 1935-40 (1996)). However, the success isfor the moment still limited. None of these alternatives has reached thestage of replacing polyoxyethylated castor oil based vehicle in theclinical application.

Another approach to overcome the hypersensitivity reactions resultingfrom polyoxyethylated castor oil can be the design of oral formulationsof paclitaxel (J. M. M. Terwogt, M. M. Malingre, J. H. Beijnen, W. W. B.Huinink, H. Rosing, F. J. Koopman, O. van Tellingen, M. Swart, and J. H.M. Schellens. Coadministration of oral cyclosporin A enables oraltherapy with paclitaxel. Clin Cancer Res. 5: 3379-84 (1999)). Oraladministration of paclitaxel would, thus, prevent the adverse effectscaused by the vehicle substance polyoxyethylated castor oil and offeradditional advantages over intravenous administration, includingelimination of the need for frequent visits to the outpatient clinic andeasier chronic administration (R. T. Dorr. Pharmacology and toxicologyof Cremophor EL diluent. Ann Pharmacother. 28: S11-4 (1994); A. J. tenTije, J. Verweij, W. J. Loos, and A. Sparreboom. Pharmacological effectsof formulation vehicles : implications for cancer chemotherapy. ClinPharmacokinet 42: 665-85 (2003)). However, preclinical studies havesuggested that paclitaxel is not significantly absorbed after oraladministration; the systemic bioavailability in humans after oralpaclitaxel administration is less than 6% (J. M. M. Terwogt, M. M.Malingre, J. H. Beijnen, W. W. B. Huinink, H. Rosing, F. J. Koopman, O.van Tellingen, M. Swart, and J. H. M. Schellens. Coadministration oforal cyclosporin A enables oral therapy with paclitaxel. Clin CancerRes. 5: 3379-84 (1999)). The explanations proposed to account for thepoor oral bioavailability of paclitaxel are multifactorial. The mostlikely explanations are its affinity for the membrane-bound drug effluxpump P-glycoprotein (P-gp), metabolization by cytochromes P450 (2C8 and3A4) and poor water solubility.(R. T. Liggins, W. L. Hunter, H. M. Burt.Solid-state characterization of paclitaxel. J. Pharm Sci. 86: 1458-63(1997); J. van Asperen, 0. van Tellingen, A. Sparreboom, A. H. Schinkel,P. Borst, W. J. Nooijen, and J. H. Beijnen. Enhanced oralbioavailability of paclitaxel in mice treated with the P-glycoproteinblocker SDZ PSC 833. Br J. Cancer. 76: 1181-3 (1997); C. D. Britten, S.D. Baker, L. J. Denis, T. Johnson, R. Drengler, L. L. Siu, K. Duchin, J.Kuhn, and E. K. Rowinsky. Oral paclitaxel and concurrent cyclosporin A:targeting clinically relevant systemic exposure to paclitaxel. ClinCancer Res. 6: 3459-68 (2000)). Moreover, the polyethoxylated castor oil(Cremophor EL) was shown to be in part responsible of the lowbioavailability and poor pharmacokinetic linearity of orallyadministered Taxol® (H. A. Bardelmeijer, M. Ouwehand, M. M. Malingre, J.H. Schellens, J. H. Beijnen, and O. van Tellingen. Entrapment byCremophor EL decreases the absorption of paclitaxel from the gut. CancerChemother Pharmacol 49: 119-125 (2002); M. M. Malingre, J. H. Schellens,O. Van Tellingen, M. Ouwehand, H. A. Bardelmeijer, H. Rosing, F. J.Koopman, M. E. Schot, W. W. Ten Bokkel Huinink, and J. H. Beijnen. Theco-solvent Cremophor EL limits absorption of orally administeredpaclitaxel in cancer patients. Br J. Cancer 85: 1472-1477 (2001)).

A number of studies have been carried out to verify in both animals andpatients if the oral bioavailability of paclitaxel could be improvedwhen the drug is administered with P-gp or cytochrome P450 inhibitors(R. T. Dorr. Pharmacology and toxicology of Cremophor EL diluent. AnnPharmacother. 28: S11-4 (1994); J. van Asperen, O. van Tellingen, A.Sparreboom, A. H. Schinkel, P. Borst, W. J. Nooijen, and J. H. Beijnen.Enhanced oral bioavailability of paclitaxel in mice treated with theP-glycoprotein blocker SDZ PSC 833. Br J. Cancer. 76: 1181-3 (1997); C.D. Britten, S. D. Baker, L. J. Denis, T. Johnson, R. Drengler, L. L.Siu, K. Duchin, J. Kuhn, and E. K. Rowinsky. Oral paclitaxel andconcurrent cyclosporin A: targeting clinically relevant systemicexposure to paclitaxel. Clin Cancer Res. 6: 3459-68 (2000)).Cyclosporine A (CsA), a well-known immunosuppressive agent, was shown tobe one of the most promising P-gp inhibitors to enhance the oralabsorption of paclitaxel (J. M. M. Terwogt, M. M. Malingre, J. H.Beijnen, W. W. B. Huinink, H. Rosing, F. J. Koopman, O. van Tellingen,M. Swart, and J. H. M. Schellens. Coadministration of oral cyclosporin Aenables oral therapy with paclitaxel. Clin Cancer Res. 5: 3379-84(1999); C. D. Britten, S. D. Baker, L. J. Denis, T. Johnson, R.Drengler, L. L. Siu, K. Duchin, J. Kuhn, and E. K. Rowinsky. Oralpaclitaxel and concurrent cyclosporin A: targeting clinically relevantsystemic exposure to paclitaxel. Clin Cancer Res. 6: 3459-68 (2000)).CsA is a registered drug and thus is more readily available for clinicalstudies. The concomitant use of cyclosporin A for oral Taxoladministration led to an increased AUC of paclitaxel (bioavailability of20%). Nevertheless, this AUC enhancement was only reached for low doses.On the contrary, at higher doses administration showed a non linearpharmacokinetic in both rodents and human. A five fold increase (from 60to 300mg/m²) of the dose in human only led to a 2 fold increase ofplasmatic AUC (M. M. Malingre, J. M. Terwogt, J. H. Beijnen, H. Rosing,F. J. Koopman, O. van Tellingen, K. Duchin, W. W. Huinink, M. Swart, J.Lieverst, and J. H. Schellens. Phase I and pharmacokinetic study of oralpaclitaxel. J Clin Oncol 18: 2468-2475. (2000)). This non lineardose-AUC relationship is a significant obstacle to the use of oralTaxol®. Moreover, oral Taxol® exhibited a poor tolerability in patientsand occasioned acute gastro-intestinal disorders such as nausea andvomiting. The formulation contains a high amount of ethanol (50%) andhigh clinically required doses lead to a significant amount of ingestedethanol. Moreover, the formulation is very bitter due to the presence ofCremophor EL. In conclusions, the oral administration of Taxol® isgreatly limited by the bad tolerability after ingestion. The non linearpharmacokinetic lead the clinician to investigate high doses of Taxol®(>300 mg/m² paclitaxel) (M. M. Malingre, J. M. Terwogt, J. H. Beijnen,H. Rosing, F. J. Koopman, O. van Tellingen, K. Duchin, W. W. Huinink, M.Swart, J. Lieverst, and J. H. Schellens. Phase I and pharmacokineticstudy of oral paclitaxel. J Clin Oncol 18: 2468-2475. (2000)). In aphase II trial of weekly oral paclitaxel plus cyclosporine in patientswith advanced non-small-cell lung cancer, interpatient variability wascalculated at 40 to 45% and intra-individual variability at 15% (C. M.Kruijtzer, J. H. Schellens, J. Mezger, M. E. Scheulen, U. Keilholz, J.H. Beijnen, H. Rosing, R. A. Mathot, S. Marcus, H. van Tinteren, and P.Baas. Phase II and pharmacologic study of weekly oral paclitaxel pluscyclosporine in patients with advanced non-small-cell lung cancer. JClin Oncol 20: 4508-16 (2002)). Those points also represent importantTaxol® limitations in the oral paclitaxel treatment.

Recently, it was reported that self-emulsifying oily formulation (SEOF)consisting of isotropic mixtures of oil and surfactants couldsignificantly improve the oral availability of poorly absorbed,hydrophobic and/or lipophilic drugs (T. Gershanik, S. Benita.Self-dispersing lipid formulations for improving oral absorption oflipophilic drugs. Eur J Pharm Biopharm. 50: 179-88 (2000)). SEOFs arecomposed of natural or synthetic oils, surfactants and one or morehydrophilic solvents and co-solvents. The principal characteristic ofSEOFs is their ability to form fine oil-in-water emulsions ormicroemulsions upon mild agitation following dilution by aqueous phases.These formulations can disperse in the gastrointestinal lumen to formmicroemulsions or fine emulsions, upon dilution with gastrointestinalfluids. In in-vivo absorption studies in non-fasting dogs, SEOFselicited at least a three-fold greater C_(max) and AUC of a lipophilicnaphthalene derivative than that of the drug in any other dosage form(N. H. Shah, M. T. Carvajal, C. I. Patel, M. H. Infeld, A. W. Malick.Self-emulsifying drug delivery systems (SEDDS) with polyglycolyzedglycerides for improving in vitro dissolution and oral absorption oflipophilic drugs. Int J Pharm. 106: 15-23 (1994)). The absorption ofontazolast in rats was significantly enhanced by all lipid-basedformulations (D. J. Hauss, S. E. Fogal, J. V. Ficorilli, C. A. Price, T.Roy, A. A. Jayaraj, and J. J. Kierns. Lipid-based delivery systems forimproving the bioavailability and lymphatic transport of a poorlywater-soluble LTB4 inhibitor. J Pharm Sci. 87: 164-9 (1998)).Microemulsions have successfully been used to improve drugsolubilization/dissolution and/or intestinal absorption of poorlyabsorbed drugs including CsA (P. P. Constantinides. Lipid microemulsionsfor improving drug dissolution and oral absorption: physical andbiopharmaceutical aspects. Pharm Res. 12: 1561-72 (1995); S. Tenjarla.Microemulsions: an overview and pharmaceutical applications. Crit RevTher Drug Carrier Syst. 16: 461-521 (1999)).

Traditional surfactants are known to entrap lyophobic drugs. Forexample, addition of Cremophor EL to the formulation of oral drugpreparations resulted in significantly diminished drug uptake andreduced circulating concentrations. The drawbacks presented by thepresence of Cremophor EL or Tween 80 in drug formulations haveinstigated extensive research to develop alternative delivery forms.Currently, several strategies are in progress to develop Tween 80 andCremophor EL-free formulations of docetaxel and paclitaxel, which arebased on pharmaceutical, chemical or biological strategies (A. J. tenTije, J. Verweij, W. J. Loos, and A. Sparreboom. Pharmacological effectsof formulation vehicles: implications for cancer chemotherapy. ClinPharmacokinet 42: 665-85 (2003); H. A. Bardelmeijer, M. Ouwehand, M. M.Malingre, J. H. Schellens, J. H. Beijnen, and O. van Tellingen.Entrapment by Cremophor EL decreases the absorption of paclitaxel fromthe gut. Cancer Chemother Pharmacol 49: 119-125 (2002)).

The rationale of a self-emulsifying oily formulation for theadministration of oral paclitaxel lies in the better solubilization andabsorption of paclitaxel and concomitant bioavailability variabilityreduction.

There is a continuing need for taxane compositions and formulationswhich provide a more efficient means of administering taxanes withoutcausing undesired side effects and which have improved stability andlonger shelf life.

An object of the instant invention is a pharmaceutical composition in aform of an anhydrous self-nanoemulsifying oily formulation comprising:

one or more therapeutic agent(s) which have low solubility in water orare water-insoluble,

vitamin E,

one co-solvent selected from propylene glycol and ethanol and mixturethereof

one surfactant selected from tyloxapol and mixture of tyloxapol and TPGSoptionally,

a bioenhancer.

In a specific aspect of the instant invention, the pH of the compositioncan be reduced to further improve the stability of the therapeuticagent. In some embodiments this is accomplished by the addition of anacidic pH adjuster which is selected from the group comprising ascorbicacid, citric acid, tartaric acid, lactic acid, oxalic acid, formic acid,benzene sulphonic acid, benzoic acid, maleic acid, glutamic acid,succinic acid, aspartic acid, diatrizoic acid, and acetic acid. Theacidifying agent may also be an inorganic acid, including, but notlimited to, hydrochloric acid, sulphuric acid, phosphoric acid, andnitric acid. An anhydrous organic acid, like anhydrous citric acid, maypreferably be used in the composition.

In another specific embodiment of the pharmaceutical composition vitaminE is from 2 to 6% (w/w) of the final composition.

According to the invention, the one or more therapeutic agent(s) isselected from the group comprising anti-fungal drugs, anti-viral drugs,antibiotic drugs, anti-inflammatory drugs, anti-cancer drugs,analgesics, antidepressants, antipsychotics, hormones, antacids,coronary vasodilators, cerebral vasodilators, psychotropics,antineoplastics, stimulants, anti-histamines, vasodilators,anti-arrythmics, anti-hypertensive drugs, vasoconstrictors,anti-migraine drugs, anti-coagulants and anti-thrombotic drugs,anti-pyretics, hypnotics, sedatives, anticonvulsants, anti-epileptics,neuromuscular drugs, drugs acting on Central Nervous System, hyper- andhypoglycemic agents, diuretics, anti-obesity drugs, anabolic drugs,anti-uricemic drugs and combinations thereof.

In a specific embodiment, the anti-cancer drug is a taxoid, preferablyselected from paclitaxel, docetaxel, their derivatives, analogs andprodrugs.

When the taxoid is paclitaxel, it is present in a relative proportionbetween 0.5 and 4% (w/w) of the final composition, preferably between1.5 and 3% (wlw).

According to one specific embodiment of the invention, preferredpharmaceutical composition for oral use comprises an emulsion includingvitamin E, D-α-tocopheryl polyethylene glycol succinate 1000 (TPGS),tyloxapol and at least, one therapeutic agent.

The relative proportions of vitamin E, TPGS and tyloxapol may berespectively 2-6, 0-60 and 5-70 (w/w) of the final composition,preferably respectively 2-6, 5-60 and 5-70(w/w) of the finalcomposition, more preferably respectively 3-5, 20-40 and 20-40%.

When the composition is used for intravenous route it contains no TPGS.

According to another specific embodiment of the invention, the relativeproportion of propylene glycol is in the range of 0-50% (w/w) of thefinal composition, preferably equal to 20% (w/w) and the relativeproportion of ethanol is in the range of 5-50% (w/w) of the finalcomposition, preferably equal to 30% (w/w).

According to the instant invention, the enhancer is one well-known fromthe man skilled in the art. It is advantageously selected from the groupcomprising cytochrome P450 2C8 inhibitors, cytochrome P450 3A4inhibitors, multidrug resistance inhibitors, Pgp inhibitors or nonspecific inhibitors.

In a specific embodiment, the enhancer is selected from cyclosporine A,its analogs and derivatives.

The compositions according to the invention may be associated with anypharmaceutical excipient to form a dosage form, which can beadministered to animals or humans via intravascular, oral,intramuscular, cutaneous and subcutaneous routes. Specifically emulsionsaccording to the invention can be given by any of the following routesamong others: intra-abdominal, intra-arterial, intra-articular,intra-capsular, intra-cervical, intra-cranial, intra-ductal,intra-dural, intra-lesional, intra-ocular, intra-locular, intra-lumbar,intra-mural, intra-operative, intra-parietal, intra-peritoneal,intra-plural, intra-pulmonary, intra-spinal, intra-thoracic,intra-tracheal, intra-tympanic, intra-uterine, intra-ventricular,intra-venous or transdermal or can be nebulised using suitable aerosolpropellants.

Self-emulsifying systems give emulsions upon dilution in aqueous media.The presence of an oily core in emulsion droplets (nonexistent inmicelles) enables to dissolve higher quantity of drugs and provides anencapsulation effect of stabilization. The presence of an oily core(droplets instead of micelles) enables to load bigger quantity of drugs(e.g. paclitaxel) within the SNEOF than traditional micelles-formingsystems such as Taxol®. Furthermore, at equivalent drug concentration,the emulsion provides a better stability of paclitaxel (degradationand/or precipitation) than micelles. The self-nanoemulsifying oilyformulations give nanoemulsions with droplet size smaller than or equalto 10 nanometers. Incorporation of drugs such as paclitaxel orcyclosporin A within the SNEOFs does not significantly alter theemulsion size nor the self-nanoemulsification process. The surfaceoffered to drug release and absorption is huge and represents more than600 m² for 1 ml of SNEOF.

The compositions according to the invention have been specificallydesigned to ensure self-nanoemulsification (viscosity, HLB) and toprovide the best drug solubilization properties. Moreover, thesurfactant choice was conditioned by the ability to release the drug soas to ensure a linear pharmacokinetic even after oral administration.

In the sense of the present invention, when the pharmacokinetic islinear, the dose of therapeutic agent is proportional to the bloodplasma level of the therapeutic agent desired.

Thus, two novel polymeric surfactants were used concomitantly with anoily carrier: tyloxapol (oxyethylated tert-octylphenol formaldehydepolymer) and alpha-tocophéryl polyethylene glycol 1000 succinate (TPGS).Their unique combination in SNEOF ensures a linear release of lyophobicdrugs and a linear pharmacokinetic even at high doses.

An other object of the invention is the use of TPGS and tyloxapol forpreparing pharmaceutical composition in the form of anhydrousself-nanoemulsifying oily formulation having linear pharmacokinetic evenat high doses after oral administration.

Another aspect of the invention is a method of treatment oftaxoïd-responsive diseases wherein an effective amount of a compositionaccording to the invention is administered to a patient in need thereof

The advantages of paclitaxel SNEOF according to the instant inventionover orally given Taxol® are:

-   -   absence of Cremophor EL,    -   better taste, less bitterness,    -   2.5 to 5 fold bigger Paclitaxel content,    -   4 to 8 fold less ethanol at equivalent dose and    -   1.25 to 2.5 fold less ingested volume.

Even low paclitaxel doses of 60 or 90 mg/m² with oral Taxol® lead to badpatient tolerability (nausea, vomiting) (C. M. Kruijtzer, H. Boot, J. H.Beijnen, H. L. Lochs, F. X. Pamis, A. S. Planting, J. M. Pelgrims, R.Williams, R. A. Mathot, H. Rosing, M. E. Schot, H. Van Tinteren, and J.H. Schellens. Weekly oral paclitaxel as first-line treatment in patientswith advanced gastric cancer. Ann Oncol 14: 197-204 (2003)). A bettertolerability after ingestion is assumed based on a significativereduction of the ethanol content and of the bitterness reduction.Furthermore, selected excipients (TPGS, tyloxapol) have a lower rodentoral DL50 than Cremophor EL. The reduced volume and reduced overalltoxicity could safely allow to administrate higher paclitaxel dose topatients than oral Taxol® formulation for which the maximal tolerateddose (MTD) was estimated at 360 mg/m² (M. M. Malingre, J. M. Terwogt, J.H. Beijnen, H. Rosing, F. J. Koopman, O. van Tellingen, K. Duchin, W. W.Huinink, M. Swart, J. Lieverst, and J. H. Schellens. Phase I andpharmacokinetic study of oral paclitaxel. J Clin Oncol 18: 2468-2475.(2000)). Moreover the dose-AUC non-linearity and pharmacokineticvariabilities of orally given Taxol® constitute the main limitations toan oral clinical practice use and security (M. M. Malingre, J. H.Beijnen, H. Rosing, F. J. Koopman, O. van Tellingen, K. Duchin, W. W.Ten Bokkel Huinink, M. Swart, J. Lieverst, and J. H. Schellens. A phaseI and pharmacokinetic study of bi-daily dosing of oral paclitaxel incombination with cyclosporin A. Cancer Chemother Pharmacol 47: 347-54(2001)).

The self-nanoemulsifying compositions according to the instant inventionmay be used for the treatment of different diseases like cancers,tumours, Kaposi's sarcoma, malignancies, uncontrolled tissue or cellularproliferation secondary to tissue injury, and any other diseaseconditions responsive to taxoids such as paclitaxel and docetaxel,and/or prodrugs and derivatives of the foregoing. Among the types ofcarcinoma which may be treated particularly effectively with oralpaclitaxel, docetaxel, other taxoids, and their prodrugs andderivatives, are hepatocellular carcinoma and liver metastases, cancersof the gastrointestinal tract, pancreas, prostate and lung, and Kaposi'ssarcoma. Examples of non-cancerous disease conditions which may beeffectively treated with these active agents administered orally inaccordance with the present invention are uncontrolled tissue orcellular proliferation secondary to tissue injury, polycystic kidneydisease, inflammatory diseases (e. g., arthritis) and malaria.

The novel compositions may be administered in any known pharmaceuticaloral dosage form. For example, the formulations may be encapsulated in asoft or hard gelatin capsule or may be administered in the form of aliquid oily preparation. Each dosage form may include, apart from theessential components of the composition conventional pharmaceuticalexcipients, diluents, sweeteners, flavouring agents, colouring agentsand any other inert ingredients regularly included in dosage formsintended for oral administration (see e. g., Remington's PharmaceuticalSciences, 17th Ed., 1985).

Precise amounts of each of the target drugs included in the oral dosageforms will vary depending on the age, weight, disease and condition ofthe patient.

Although some of the oral formulations of the invention may providetherapeutic blood levels of the taxoid active ingredient whenadministered alone, an advantageous method of the invention for treatingmammalian patients (particularly human patients) suffering fromtaxoid-responsive disease conditions is to administer the oralformulations containing the taxoid target agent concomitantly with theadministration of at least one dose of an oral bioavailability enhancingagent. This bioenhancer can be concomitantly formulated in the SNEOF oradministered separately. Another advantageous method of the inventionfor treating mammalian patients is to administer the oral formulationscontaining the taxoid target agent concomitantly or separately withanother antitumor agent like carboplatinum and the like.

The preferred embodiment of the method of the invention for oraladministration to humans of paclitaxel, its derivatives, analogs andprodrugs, and other taxoids comprises the oral administration of an oralabsorption or bioavailability enhancing agent to a human patientsimultaneously with, or prior to, or both simultaneously with and priorto the oral administration to increase the quantity of absorption of theintact target agent into the bloodstream.

Different advantages of the present invention will be readilyappreciated with the following figures, tables and examples.

FIG. 1 illustrates the pharmacokinetic profiles of 10 mg/kgintravenously administered paclitaxel with or without 10 mg/kgcyclosporin A oral pre-treatment.

FIG. 2 illustrates the pharmacokinetic profiles of oral 10 mg/kgpaclitaxel formulations according to the invention with or without 10mg/kg cyclosporin A oral pre-treatment and compared to paclitaxel alone.

FIG. 3 illustrates the pharmacokinetic profiles of oral 10, 30 and 60mg/kg paclitaxel formulations according to the invention without 10mg/kg cyclosporin A oral pre-treatment in P-gp knock out mice.

FIG. 4 illustrates the pharmacokinetic profiles of oral 10, 30 and 60mg/kg paclitaxel formulations according to the invention with 10 mg/kgcyclosporin A oral pre-treatment in wild type mice.

EXAMPLE 1 Characterization of Paclitazel Emulsions Following 1:10Dilution of Sneofs with Water

1. Materials and Methods

1.1. Materials

Paclitaxel (MW 853) with 99.34% (w/w) purity (HPLC) was purchased fromFarmachem (Lugano, Switzerland). Vitamin E and tyloxapol were boughtfrom Sigma (St. Louis, Mo., USA). D-α-tocopheryl polyethylene glycolsuccinate 1000 (TPGS) was a gift from Eastman Chemical (Kingsport,Tenn., USA). Ethanol was bought from SDS (Peypin, France). All solventswere HPLC grade

1.2. Methods

Preparation of Paclitaxel SNEOFs

SNEOFs were firstly prepared by successive addition and mixing of eachexcipient. When the oily carrier is clear and homogeneous, paclitaxel isadded and quickly dissolved under mild agitation. For combinedformulations, Cyclosporin A is lastly added and quickly dissolved undermild agitation in the SNEOF. In the same manner, pH-lowered formulationsare obtained by addition of 0.01 or 0.02 % anhydrous citric acid in thepreviously prepared paclitaxel-containing SNEOF. Its dissolution is slowand requires agitation.

Paclitaxel emulsion may be formed by dilution of SNEOFs with distilledwater.

Examples of Paclitaxel SNEOFs

The following compositions have been prepared according to theabove-disclosed method. Components % (w, w) Paclitaxel 1.5 Vitamin E 5TPGS 31.75 Ethanol anhydrous, absolute 30 Tyloxapol 31.75 Paclitaxel 3Vitamin E 5 TPGS 31 Ethanol anhydrous, absolute 30 Tyloxapol 31Paclitaxel 1.5 Vitamin E 5 Ethanol anhydrous, absolute 30 Tyloxapol 63.5Examples of Combined Formulations

The following compositions have been prepared according to theabove-disclosed method. Components % (w, w) Paclitaxel 1.5 Vitamin E 5TPGS 31 Ethanol anhydrous, absolute 30 Tyloxapol 31 Cyclosporin A 1.5Paclitaxel 3 Vitamin E 5 TPGS 30.25 Ethanol anhydrous, absolute 30Tyloxapol 30.25 Cyclosporin A 1.5 Paclitaxel 1.5 Vitamin E 5 Ethanolanhydrous, absolute 30 Tyloxapol 62 Cyclosporin A 1.5Examples of pH-Lowered Formulations

The following compositions have been prepared according to theabove-disclosed method. Components % (w, w) Paclitaxel 1.5 Vitamin E 5TPGS 31.75 Ethanol anhydrous, absolute 30 Tyloxapol 31.75 Citric acid0.01Droplet Size

Emulsions were formed following 1:10 dilution of paclitaxel SNEOF withdistilled water. The droplet size of the resulting emulsions wasdetermined by the PCS method using a Nanosizer (Malvern, UK)

Stability Study

SNEOFs containing 1.5 and 3% (w/w) paclitaxel were prepared. A combinedform of 1.5% paclitaxel plus 1.5% cyclosporin A was also prepared. Thechemical stability of paclitaxel in SNEOFs was monitored using ananalytical HPLC method (M. Andreeva, P. D. ledmann, L. Binder, V. W.Armstrong, H. Meden, M. Binder, M. Oellerich. A simple and reliablereversed-phase high-performance liquid chromatographic procedure fordetermination of paclitaxel (taxol) in human serum. Ther Drug Monit. 19:327-32 (1997); A. Sharma, W. D. Conway, R. M. Straubinger.Reversed-phase high-performance liquid chromatographic determination oftaxol in mouse plasma. J Chromatogr B Biomed Appl. 655: 315-9 (1994)).

2. Results

2.1. Physicochemical Characterization

Droplet Size

Following 1:10 dilution of paclitaxel SNEOFs (1.5% and 3% w/w) indistilled water, the droplet size of the resulting nanoemulsions was inaverage equal to 10±4.0 nm with a low Polydispersity index (<0.15).

2.2. Stability Study

The preliminary chemical stability studies indicated that paclitaxel inthe SNEOFs was stable at 4, 25 and 40° C. The drug content in SNEOFs at4, 25 and 40° C. did not change over three months.

EXAMPLE 2 Pharmacokinetic Study of Oral Paclitaxel Emulsions AfterCyclosporin a Oral Pretreatment

1. Materials and Methods

1.1. Materials

1.1.1. Animals

Strains: FVB Wild-type mice

Source: Breeding stocks of the animal facility of the Netherlands KancerInstitute (NKI)

Age: 8-14 weeks

Body weight: 18-30 gram

Gender: Female

Housing: Animal Department of the NKI

1.1.2. Drug

Oral Formulation: Cyclosporin A (Sandimmun®)

Source: Novartis

Vehicle: Cremophor EL: Ethanol (65:35, v/v)

Concentration: 50 mg/mL

Route: Oral (p.o.)

Dose: 10 mg/kg

Oral Taxol: Paclitaxel (Taxol®)

Source: Bristol-Myers Squibb

Vehicle: Cremophor EL: Ethanol (1:1; v/v)

Concentration: 6 mg/mL and 2 mg/mL after 1:3 dilution in water forinjection (WFI)

Route: Oral (p.o.)

Dose: 10 mg/kg

Oral Formulation 1: Paclitaxel SNEOF

Source: Novagali SAS

Vehicle: Tyloxapol/TPGS/Ethanol/Vitamin E

Concentration: 15 mg/mL for SEOF and 1.5 mg/mL after 1:10 dilution inWFI

Route: Oral (p.o)

Dose: 10 mg/kg

Oral formulation 1 composition according to the invention Components %(w, w) Paclitaxel 1.5 Vitamin E 5 TPGS 31.75 Ethanol anhydrous, absolute30 Tyloxapol 31.75

Oral Formulation 2 according to the invention: Paclitaxel/Cyclosporin ASNEOF

Source: Novagali SAS

Vehicle: Tyloxapol/TPGS/Ethanol/Vitamin E

Concentration: Paclitaxel: 15 mg/mL for SEOF and 1.5 mg/mL after 1:10dilution in WFI

Cyclosporin A: 15 mg/mL for SEOF and 1.5 mg/mL after 1:10 dilution

Route: Oral (p.o)

Dose: 10 mg/kg paclitaxel and 10 mg/kg cyclosporin A Oral formulation 2composition Components % (w, w) Paclitaxel 1.5 Vitamin E 5 TPGS 31Ethanol anhydrous, absolute 30 Tyloxapol 31 Cyclosporin A 1.5I.V. Formulation: Paclitaxel

Source: Paclitaxel (pure compound) supplied by Novagali

Vehicle: Polysorbate 80: Ethanol (1:1, v/v)

Concentration: 6 mg/mL, 1.5 mg/ml after 1:4 dilution in saline

Route: intravenous (i.v.)

Dose: 10 mg/kg

1.1.3. Preparation of Drug Solutions for Oral Administration

Cyclosporin A (Sandimmun®) was diluted 1:25 in water for injection toyield a final concentration of 2 mg/mL. A volume of 5 μL per gram bodyweight was administered to the animals resulting in a dose of 10 mg/kgof cyclosporin A.

Paclitaxel self-nanoemulsifying oily formulations (SNEOF) n°1 and 2 werediluted 1:10 in WFI to give a microemulsion. A volume of 6.67 μL pergram (body weight) was administered to the animals resulting in a doseof 10 mg/kg of paclitaxel.

Paclitaxel in Cremophor EL: ethanol (Taxol®) was diluted with water forinjection to a final concentration of 1.5 mg/ml (1:4 dilution). A volumeof 6.67 μL per gram (body weight) was administered resulting in a doselevel of 10 mg/kg of paclitaxel.

1.1.4. Preparation of Drug Solutions for Intravenous Administrations

The stock solution of paclitaxel in Polysorbate 80:ethanol (1:1; v/v)was diluted 1:4 with saline to achieve a final concentration of 1.5mg/mL. A volume of 6.67 μL per gram (body weight) was administered tothe animals, resulting in a dose level of 10 mg/kg of paclitaxel.

1.2. Study Set Up CsA oral Paclitaxel Animal Blood sampling CohortPre-treatment vehicle number times 1 10 mg/kg TAXOL 0.6% 20 1, 2, 4 & 8h 2 10 mg/kg SNEOF 1.5% 20 1, 2, 4 & 8 h 3 None SNEOF 1.5% + 20 1, 2, 4& 8 h 1.5% CsA 4 None PS 80: EtOH 24 5 & 30 min, 1, 0.6% IV 2, 4 & 8 h 510 mg/kg PS 80: EtOH 24 5 & 30 min, 1, 0.6% IV 2, 4 & 8 h

Each cohort through 1 to 3 consisted of 20 Wild-type mice and cohorts.Cohort 1 received paclitaxel in the conventional formulation ofCremophor EL and ethanol at a dose of 10 mg/kg.

Cohorts 2 received a paclitaxel microemulsion formulation according tothe invention at a dose of 10 mg/kg of paclitaxel.

Cohorts 1 and 2 received a pre-treatment of 10 mg/kg cyclosporin A, 30minutes prior to paclitaxel administration.

Cohort 3 received both cyclosporin and paclitaxel in a microemulsionformulation according to the instant invention at dose levels of 10mg/kg of paclitaxel and 10 mg/kg of cyclosporin A.

Cohorts 4 and 5 are reference groups for the calculation of thebioavailability and consisted of 24 animals per cohort. They receivedpaclitaxel at a dose of 10 mg/kg by intravenous injection in the tailvein following oral cyclosporin A or cyclosporin A vehicle.

Oral drug administrations was done by stainless steel gavage using aglass syringe (250 μl: Hamilton luer tip) or a disposable polypropylenesyringe (1 ml). The gavage was inserted via the oesophagus into thestomach. Cyclosporin A was administered orally 18±2 min prior to oralpaclitaxel. At times 1, 2, 4 and 8 h after oral paclitaxeladministration, animals (n=5 per time point per group) wereanaesthetized with metofane and blood was sampled by cardiac puncture.

Intravenous drug administrations were done by injection into the tailvein using a disposable polypropylene syringe (300 μl) provided withfixed 29 g needle. Cyclosporin A was administered 30±5 min prior tointravenous paclitaxel. At times 5, 30 min, 1, 2, 4 and 8 h afterintravenous paclitaxel administration, animals (n=4 per time point pergroup) were anaesthetized with metofane and blood was sampled by cardiacpuncture.

1.2.1. Sample Collection, Handling and Storage

At the specified times, animals were anaesthetized using metofane. Afterfixation on their back, with their chest in an upright position bloodwas collected by cardiac puncture using a 1 mL polypropylene syringefitted with 25 g needle. Blood was transferred immediately into tubescontaining potassium EDTA as anticoagulant and mixed by inversion. Bloodsamples were centrifuged vial for 5 min at 4000 g. The supernatantplasma fraction was transferred to a clean tube with appropriate labeland stored at −20° C. until analyses.

1.2.2. Number of Animals/Samples

Based on previous experience, 5 animals per time point was sufficient toaccurately determine the AUC of the plasma concentration-time curvesafter oral dosing, whereas 4 animals per time point sufficed forintravenous dose groups. Plasma samples were obtained from all animals.

1.2.3. Analytical Method

Analyses of paclitaxel levels in the plasma samples used a validatedHPLC-UV methodology (Sparreboom, A., van Tellingen, O., Nooijen, W. J.,and Beijnen, J. H. Determination of paclitaxel and metabolites in mouseplasma, tissues, urine and faeces by semi-automated reversed-phasehigh-performance liquid chromatography. J Chromatogr B Biomed Appl, 664:383-391, 1995.).

1.2.4. Assay Performance Control

Test samples were analysed for paclitaxel in singular within ananalytical batch, consisting of a set of calibration standards and QCsamples. Per series of 40 test samples at least 3 QC samples containingpaclitaxel at concentrations over the expected range were analysed induplicate. Results of batch analysed were accepted if:

the correlation coefficient (r) of the calibration curve is higher than0.98.

at least 4 of the 6 QC samples are within ±20% of their respectivenominal values; 2 of the 6 QC samples (not both at the sameconcentration) may be outside the ±20% of their respective nominalvalues.

1.2.5. Data Reprocessing

Chromatographic data acquisition and processing were done using aChromeleon (v.6) chromatography data station. Calibration lines werefitted by weighed least squares regression analysis using the reciprocalof the squared concentration as the weight factor.

Plasma concentration data were reported. Plasma concentrations versustime curves were fitted using the MEDI\WARE software package (version3.0) and pharnacokinetic parameters: Area under the plasmaconcentration-time curve (AUC), Maximum plasma level (C_(max)),Elimination half-life (t½) and Biological availability (F) werecalculated.

2. Results

They are illustrated in FIGS. 1 and 2 and in the following table.

Paclitaxel bioavailability of 10 mg/kg paclitaxel oral formulations withor without 10 mg/kg oral cyclosporin A pre-medication CsA oral AUCBioavail- Pre- Paclitaxel (mean ± SE) ability Cohort treatment vehicle(ng/ml · h) (%) 1 10 mg/kg TAXOL 0.6% 2984 ± 173 21.3 2 10 mg/kg SNEOF1.5% 2670 ± 249 19.1 3 None SNEOF 1.5% + 2772 ± 344 19.8 1.5% CsA 4 NonePS 80: EtOH 5961 ± 374 100 0.6% IV 5 10 mg/kg PS 80: EtOH 14006 ± 725 100 0.6% IV3. Conclusion

Cyclosporin A is known to be an oral taxanes bioenhancer. Oralpre-treatment with cyclosporin A lead to a significant increase ofintravenously administered paclitaxel AUC from 5961±374 to 14006±725(2.35 fold). Oral pre-treatment with cyclosporin A also lead to asignificant increase of orally administered paclitaxel AUC (data notshown).

At the dose of 10 mg/kg of paclitaxel plus 10 mg/kg of Cyclosporin Aorally administered to wild type mice, both the SNEOF and the Taxol®formulation exhibited an equivalent bioavailability of approximately20%.

The paclitaxel and cyclosporin A-containing SNEOF also lead to a 20%bioavailability of paclitaxel. This formulation illustrates thepossibility to use a combined oral dosage form (SNEOF) of the drug and abioenhancer.

EXAMPLE 3 Dose Versus Plasma—Auc Pharmacokinetic Study of OralPaclitaxel Emulsions After Cyclosporin a Oral Pre-Treatmnet in Wild TypeMice or without Cyclosporin Oral Pre-Treatment in P-Glycoprotein KnockOut Mice

1. Materials and Methods

1.1. Materials

1.1.1. Animals

Strains: Mdr1 a/b KO mice and FVB Wild type Mice

Source: Breeding stocks of the animal facility of the NKI

Age: 8-14 weeks

Body weight: 18-30 gram

Gender: Female

Housing: Animal Department of the NKI

1.1.2. Drug

Oral Formulation: Paclitaxel SNEOF

Source: Novagali Pharma SA

Vehicle: Tyloxapol/TPGS/Ethanol/Vitamin E

Concentration: 15 mg/mL paclitaxel

Route: Oral (p.o)

Dose: 10, 30 and 60 mg/kg Composition of paclitaxel SNEOF Components %(w, w) Paclitaxel 1.5 Vitamin E 5 TPGS 31.75 Ethanol anhydrous, absolute30 Tyloxapol 31.75I.V. Formulation: Paclitaxel

Source: Paclitaxel (pure compound) supplied by Novagali

Vehicle: Polysorbate 80: Ethanol (1:1, v/v)

Concentration: 6 mg/mL, 1.5 mg/ml after 1:4 dilution in WFI

Route: intravenous (i.v.)

Dose: 10 mg/kg

1.1.3. Preparation of Drug Solutions for Oral and IntravenousAdministration

Paclitaxel self-nanoemulsifying oily formulations (SNEOF) were diluted1:10 in WFI to give a microemulsion containing 1.5 mg/ml of paclitaxel.A volume of 6.67 μL per gram (body weight) was administered orally tothe animals resulting in a dose of 10 mg/kg of paclitaxel.

Paclitaxel self-nanoemulsifying oily formulations (SNEOF) were diluted1:10 in WFI to give a microemulsion containing 1.5 mg/ml of paclitaxel.A volume of 20 μL per gram (body weight) was administered orally to theanimals resulting in a dose of 30 mg/kg of paclitaxel.

Paclitaxel self-nanoemulsifying oily formulations (SNEOF) were diluted1:10 in WFI to give a microemulsion containing 1.5 mg/ml of paclitaxel.A volume of 40 μL per gram (body weight) was administered orally to theanimals resulting in a dose of 30 mg/kg of paclitaxel.

The stock solution of paclitaxel in Polysorbate 80:ethanol (1:1; v/v)was diluted 1:4 with saline to achieve a final concentration of 1.5mg/mL. A volume of 6.67 μL per gram (body weight) was administeredintravenously to the animals, resulting in a dose level of 10 mg/kg ofpaclitaxel.

1.2. Study Set Up

1.2.1. Plasma Pharmacokinetic Study

Cohorts 12, 13 and 14, received paclitaxel in formulation SNEOF at doselevels of 10, 30 and 60 mg/kg, respectively to investigate the linearityof dose versus plasma AUC relationships of paclitaxel when this drug wasgiven as single agent to P glycoprotein knockout mice, whereas cohorts15, 16 and 17 received paclitaxel in formulation SNEOF at dose levels of10, 30 and 60 mg/kg, respectively to investigate the linearity of doseversus plasma AUC relationships of paclitaxel when this drug was givenafter a Cyclosporin A pre-treatment to Wild type Mice. Each cohortconsisted of 20 mice.

1.2.2. Sample Collection, Handling and Storage

Plasma Pharmacokinetic Study

At the specified times, animals were anaesthetized using metofane. Afterfixation on their back, with their chest in an upright position bloodwas collected by cardiac puncture using a 1 mL polypropylene syringefitted with 25 g needle. Blood was transferred immediately into tubescontaining potassium EDTA as anticoagulant and mixed by inversion. Bloodsamples were centrifuged vial for 5 min at 4000 g. The supernatantplasma fraction was transferred to a clean tube with appropriate labeland stored at −20° C. until analyses.

5 animals per time point were used to determine the AUC of the plasmaconcentration-time curves after oral dosing, whereas 4 animals per timepoint were used for intravenous dose groups. Plasma samples forsubsequent analysis were obtained from all animals.

1.3. Analytical Method

Analyses of paclitaxel levels in the plasma samples were a validatedHPLC-UV methodology (Sparreboom, A., van Tellingen, O., Nooijen, W. J.,and Beijnen, J. H. Determination of paclitaxel and metabolites in mouseplasma, tissues, urine and faeces by semi-automated reversed-phasehigh-performance liquid chromatography. J. Chromatogr B. Biomed AppI.,664: 383-391, 1995).

1.3.1. Assay Performance Control

Test samples were analysed for paclitaxel in singular within ananalytical batch, consisting of a set of calibration standards and QCsamples. Per series of 60 test samples at least 3, QC samples containingpaclitaxel at concentrations over the expected range were analysed induplicate. Results of batch analysed were accepted if:

-   -   the correlation coefficient (r) of the calibration curve is        higher than 0.98.    -   at least 4 of the 6 QC samples are within ±20% of their        respective nominal values; 2 of the 6 QC samples (not both at        the same concentration) may be outside the ±20% of their        respective nominal values.        1.3.2. Data Reprocessing

Chromatographic data acquisition and processing were done using aChromeleon (v.6) chromatography data station. Calibration lines werefitted by weighed least squares regression analysis.

Plasma concentration data were reported. Plasma concentrations versustime curves were fitted using the MEDI\WARE software package (version3.0) and pharmacokinetic parameters: Area under the plasmaconcentration-time curve (AUC), Maximum plasma level (C_(max)),Elimination half-life (t½) and Biological availability (F) werecalculated.

2. Results

Results are given in the following table and in FIG. 3 and 4.

Paclitaxel AUC and Bioavailability of oral formulations withoutCyclosporine A pre-treatment in Pgp Knock Out Mice and after CyclosporinPre-treatment in Wild type mice AUC Co- Geno- Formu- (mean ± SE)Bioavail- hort CsA type lation Route ng/ml* h ability % 12 No Mdr1 SNEOFp.o. 1935 ± 170 31.5 ± 3.1 a/b KO (10 mg/kg) 13 No Mdr1 SNEOF p.o. 7110± 382 38.6 ± 2.8 a/b KO (30 mg/kg) 14 No Mdr1 SNEOF p.o. 11229 ± 113030.4 ± 1.5 a/b KO (60 mg/kg) 15 Yes Wild SNEOF p.o. 2040 ± 174 14.8 ±1.5 type (10 mg/kg) 16 Yes Wild SNEOF p.o. 3522 ± 322  8.5 ± 0.9 type(30 mg/kg) 17 Yes Wild SNEOF p.o. 5916 ± 765  7.2 ± 1.0 type (60 mg/kg)18 No Mdr1 Poly- i.v. 6147 ± 291 a/b KO sorbate 80: EtOH3. Conclusion

As it can be observed from the Table the mean oral bioavailability ofpaclitaxel in SNEOF in P glycoprotein knockout mice ranges from 30.4 to38.6% and the oral bioavailability appears to be linear over the testeddose range as the differences in mean oral bioavailability are notstatistically different.

The AUC after 10 mg/kg of paclitaxel in wild-type mice with cyclosporineA was similar as in knockout mice receiving the same dose. Since theconcomitant administration of cyclosporine A also increased AUC ofpaclitaxel given i.v. (e.g. by inhibition of (metabolic) elimination),the oral bioavailability in wild-type mice was lower. Moreover, the oralbioavailability in wild-type animals was not linear but decreasedsignificantly with dose. In this study, the dose level of cyclosporin Awas kept constant at 10 mg/kg and it is le that the concentration ofthis competitive P gp inhibitor is insufficient using higher dose levelsof paclitaxel.

The time that the peak plasma level is reached (Tmax) increases athigher dose levels, which suggests that there may be an effect onstomach emptying and/or intestinal transit speed in mice.

1. A pharmaceutical composition in a form of an anhydrousself-nanoemulsifying oily formulation comprising: one or moretherapeutic agent(s) which have low solubility in water or arewater-insoluble, vitamin E, one co-solvent selected from propyleneglycol and ethanol and mixture thereof one surfactant selected fromtyloxapol and from mixture of tyloxapol and TPGS, and optionally, abioenhancer.
 2. A pharmaceutical composition according to claim 1further comprising an acidic pH adjuster.
 3. A pharmaceuticalcomposition according to anyone of claims 1 to 2, wherein vitamin E isfrom 2 to 6% (w/w) of the final composition.
 4. A pharmaceuticalcomposition according to anyone of claims 1 to 3, wherein the one ormore therapeutic agent(s) is selected from the group comprisinganti-fungal drugs, anti-viral drugs, antibiotic drugs, anti-inflammatorydrugs, anti-cancer drugs, analgesics, antidepressants, antipsychotics,hormones, antacids, coronary vasodilators, cerebral vasodilators,psychotropics, antineoplastics, stimulants, anti-histamines,vasodilators, anti-arrythmics, anti-hypertensive drugs,vasoconstrictors, anti-migraine drugs, anti-coagulants andanti-thrombotic drugs, anti-pyretics, hypnotics, sedatives,anticonvulsants, anti-epileptics, neuromuscular drugs, drugs acting onCentral Nervous System, hyper- and hypoglycemic agents, diuretics,anti-obesity drugs, anabolic drugs, anti-uricemic drugs,immunosuppressant drugs and combinations thereof.
 5. A pharmaceuticalcomposition according to anyone of claims 1 to 4, wherein the one ormore therapeutic agent(s) is selected from the group comprisinganti-cancer drugs, antineoplastic drugs and combinations thereof.
 6. Apharmaceutical composition according to anyone of claims 1 to 5, whereinthe anti-cancer drug is a taxoid, preferably selected from paclitaxel,docetaxel, their derivatives, analogs and prodrugs.
 7. A pharmaceuticalcomposition according to anyone of claims 1 to 6, wherein the taxoid ispaclitaxel in a relative proportion between 0.5 and 4% (w/w) of thefinal composition, preferably between 1.5 and 3% (w/w).
 8. Apharmaceutical composition according to anyone of claims 1 to 7, whereinthe relative proportions of vitamin E, TPGS and tyloxapol arerespectively 2-6, 0-60 and 5-70 (w/w) of the final composition,preferably respectively 2-6, 5-60 and 5-70 (w/w) of the finalcomposition, more preferably respectively 3-5, 20-40 and 20-40%.
 9. Apharmaceutical composition according to anyone of claim 1 to 8 whereinthe relative proportion of propylene glycol is in the range of 0-50%(w/w) of the final composition, preferably equal to 20% (w/w) and therelative proportion of ethanol is in the range of 5-50% (w/w) of thefinal composition, preferably equal to 30% (w/w).
 10. A pharmaceuticalcomposition according to anyone of claims 1 to 9, wherein the enhanceris selected from the group comprising cytochrome P450 2C8 inhibitors,cytochrome P450 3A4 inhibitors, multidrug resistance inhibitors, Pgpinhibitors or non specific inhibitors.
 11. A pharmaceutical compositionaccording to claim 10, wherein the enhancer is cyclosporine A, itsanalogs and derivatives.
 12. A pharmaceutical composition according toanyone of claims 2 to 11, wherein the acidic pH adjuster is anhydrouscitric acid.
 13. A pharmaceutical dosage form comprising an anhydrousself-nanoemulsifying oily formulation composition according to anyone ofclaims 1 to 12 associated to suitable pharmaceutical excipients.
 14. Apharmaceutical dosage form according to claim 13, which is suitable forthe oral route.
 15. A pharmaceutical dosage form according to claim 14wherein the composition is encapsulated in a soft or hard gelatincapsule or is a liquid oily preparation.
 16. A pharmaceutical dosageform according to claim 13, which is suitable for the intravenous route.17. Use of an anhydrous self-nanoemulsifying oily formulation accordingto anyone of claims 1 to 12 for the manufacture of a medicament usefulin the treatment of taxoid-responsive diseases.
 18. Use according toclaim 17 for administration to patients receiving simultaneously with,concomitantly or prior to, bioavailability enhancing agent and/oranother antitumor agent.
 19. Use of an anhydrous self-nanoemulsifyingoily formulation according to anyone of claims 1 to 12 for themanufacture of a medicament wherein the dose of the therapeutic agentadministered is linearly proportional to the blood plasma level of thetherapeutic agent desired.
 20. Use of tyloxapol and of mixture oftyloxapol and TPGS, for preparing pharmaceutical composition in the formof anhydrous self-nanoemulsifying oily formulation suitable forpreparing a medicament wherein the dose of the therapeutic agentadministered is linearly proportional to the blood plasma level of thetherapeutic agent desired.
 21. Method of treatment of taxoïd-responsivediseases wherein an effective amount of a composition according to claim1 is administered to a patient in the need thereof.